Anti-proliferative activity of biochanin A in human osteosarcoma cells via mitochondrial-involved apoptosis
Anti-proliferative activity of biochanin A in human osteosarcoma cells via mitochondrial-involved apoptosis
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DOI：
10.1016/j.fct.2017.12.062
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发表时间：
2018-02-01
影响因子：
4.3
通讯作者：
Su, Shu-Jem
中科院分区：
文献类型：
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作者：
Hsu, Yen-Nien;Shyu, Huey-Wen;Su, Shu-Jem
鹰嘴豆芽素A是红三叶草中的主要成分，也是一种有效的抗癌化学预防剂。然而，鹰嘴豆芽素A对人骨肉瘤细胞的影响从未被阐明。本研究探讨鹰嘴豆芽素A对骨肉瘤细胞的抗增殖作用。结果表明鹰嘴豆芽素A以剂量依赖性方式抑制细胞生长和集落形成，对正常细胞的毒性最小。阿霉素和鹰嘴豆芽素A联合应用可协同抑制骨肉瘤细胞的生长。鹰嘴豆芽素A通过诱导细胞凋亡的细胞毒性作用，如通过凋亡小体的形成、磷脂酰丝氨酸的外化、亚G（1）期细胞的积累、半胱天冬酶3激活和PARP裂解所证明的。细胞凋亡与线粒体膜电位的丧失、细胞色素c的释放、半胱天冬酶9的活化、Bax表达的增加以及Bcl-2和Bcl-X-L表达的减少相关。用半胱天冬酶-9特异性抑制剂（Z-LEHD-FMK）预处理部分地减弱了细胞死亡，表明内在线粒体凋亡级联的参与。然而，用JNK抑制剂SP 600125、MEK抑制剂PD-98059和p38 MAPK抑制剂SB 203580或抗氧化剂维生素E、N-乙酰半胱氨酸和谷胱甘肽预处理未能防止鹰嘴豆芽素A诱导的细胞死亡。我们的研究结果表明，鹰嘴豆芽素A抑制细胞生长，诱导骨肉瘤细胞凋亡，触发激活的内在线粒体途径和caspase-9和-3，并增加Bax：Bcl-2/Bcl-X-L的比例。
Biochanin A is a major isoflavone in red clover and a potent chemopreventive agent against cancer. However, the effects of biochanin A on human osteosarcoma cells have never been clarified. This study investigated the anti proliferative potential of biochanin A in osteosarcoma cells. The results indicate that biochanin A inhibited cell growth and colony formation in a dose-dependent manner with a minimal toxicity to normal cells. The combination of doxorubicin and biochanin A could synergistically inhibit osteosarcoma cell growth. The cytotoxic effect of biochanin A via the induction of apoptosis as evidenced by formation of apoptotic bodies, externalization of phosphatidylserine, accumulation of sub-G(1) phase cells, caspase 3 activation, and cleavage of PARP. Apoptosis was associated with loss of the mitochondrial membrane potential, release of cytochrome c, caspase 9 activation, increased Bax expression, and reduced Bcl-2 and Bcl- X-L expression. Pre-treatment with a caspase-9 specific inhibitor (Z-LEHD-FMK) partially attenuated cell death, suggesting involvement of the intrinsic mitochondrial apoptotic cascade. However, pre-treatment with the JNK inhibitor SP600125, the MEK inhibitor PD-98059, and the p38 MAPK inhibitor SB203580 or the antioxidants vitamin E, N-acetylcysteine, and glutathione failed to prevent biochanin A-induced cell death. Our results suggest that biochanin A inhibits cell growth and induces apoptosis in osteosarcoma cells by triggering activation of the intrinsic mitochondrial pathway and caspase-9 and -3 and increasing the Bax: Bcl-2/Bcl-X-L ratio.